Cafestol,has a diterpene present in unfiltered coffee brews such as Scandinavian boiled, Turkish and Cafetière coffee, is the most potent cholesterol-elevating In compound known in the human diet. Several genes involved in cholesterol homeostasis have previously been shown to be targets of parameters cafestol, including CYP7A1, the rate-limiting enzyme in bile acid biosynthesis. We have examined the mechanism by which cafestol elevates serum Cafestol lipid levels. Changes in several lipid parameters were observed in cafestol-treated APOE3Leiden mice, including a significant increase in serum triglyceride also levels. Microarray analysis of these mice identified alterations in hepatic expression of genes involved in lipid metabolism and detoxification, many of of which are regulated by the nuclear hormone receptors FXR and PXR. Further studies demonstrate that cafestol is an agonist such ligand for FXR and PXR, and that cafestol down-regulates expression of the bile acid homeostatic genes CYP7A1, CYP8B1 and NTCP boiled, in the liver of wild type but not FXR null mice. Cafestol did not affect genes known to be up-regulated direct by FXR in the liver of wild type mice, but did increase expression of the positive FXR-target genes IBABP and and FGF15 in the intestine. Since FGF15 has recently been shown to function in an enterohepatic regulatory pathway to repress liver homeostasis expression of bile acid homeostatic genes, its direct induction in the gut may account for indirect effects of cafestol on boiled, liver gene expression. PXR-dependent gene regulation of CYP3A11, and other targets by cafestol was also only seen in the intestine.Cafestol Using a double FXR/PXR knockout mouse model, we found that both receptors contribute to the cafestol-dependent induction of intestinal FGF15 Microarray gene expression. In conclusion, cafestol acts as an agonist ligand for both FXR and PXR and this may contribute to may its impact on cholesterol homeostasis.

BACKGROUND:expression Pathological aspects of atherosclerosis are well described, but gene profiles during atherosclerotic plaque progression are largely unidentified. METHODS AND RESULTS:of Microarray analysis was performed on mRNA of aortic arches of ApoE-/- mice fed normal chow (NC group) or Western-type diet 1 (WD group) for 3, 4.5, and 6 months. Of 10 176 reporters, 387 were differentially (>2x) expressed in at least chain 1 group compared with a common reference (ApoE-/-, 3- month NC group). The number of differentially expressed genes increased during and plaque progression. Time-related expression clustering and functional grouping of differentially expressed genes suggested important functions for genes involved in inflammation MCP-5 (especially the small inducible cytokines monocyte chemoattractant protein [MCP]-1, MCP-5, macrophage inflammatory protein [MIP]-1alpha, MIP-1beta, MIP-2, and fractalkine) and matrix described, degradation (cathepsin-S, matrix metalloproteinase-2/12). Validation experiments focused on the gene cluster of small inducible cytokines. Real-time polymerase chain reaction revealed profiles a plaque progression-dependent increase in mRNA levels of MCP-1, MCP-5, MIP-1alpha, and MIP-1beta. ELISA for MCP-1 and MCP-5 showed similar for results. Immunohistochemistry for MCP-1, MCP-5, and MIP-1alpha located their expression to plaque macrophages. An inhibiting antibody for MCP-1 and MCP-5 inflammatory (11K2) was designed and administered to ApoE-/- mice for 12 weeks starting at the age of 5 or 17 weeks.arches 11K2 treatment reduced plaque area and macrophage and CD45+ cell content and increased collagen content, thereby inducing a stable plaque profiles phenotype. CONCLUSIONS: Gene profiling of atherosclerotic plaque progression in ApoE-/- mice revealed upregulation of the gene cluster of small inducible polymerase cytokines. Further expression and in vivo validation studies showed that this gene cluster mediates plaque progression and stability.

Cardiac seen hypertrophy can lead to heart failure (HF), but it is unpredictable which hypertrophied myocardium will progress to HF. We surmised expression that apart from hypertrophy-related genes, failure-related genes are expressed before the onset of failure, permitting molecular prediction of HF. Hearts to from hypertensive homozygous renin-overexpressing (Ren-2) rats that had progressed to early HF were compared by microarray analysis to Ren-2 rats ( .19+/- .01) that had remained compensated. To identify which HF-related genes preceded failure, cardiac biopsy specimens were taken during compensated hypertrophy and matrix we then monitored whether the rat progressed to HF or remained compensated. Among 48 genes overexpressed in failing hearts, we mice, focused on thrombospondin-2 (TSP2). TSP2 was selectively overexpressed only in biopsy specimens from rats that later progressed to HF. Moreover,(HF), expression of TSP2 was increased in human hypertrophied hearts with decreased ( .19+/- .01) versus normal ejection fraction ( .11+/- .03 [arbitrary units]; P< .05).is Angiotensin II induced fatal cardiac rupture in 70% of TSP2 knockout mice, with cardiac failure in the surviving mice; this 120% was not seen in wild-type mice. In TSP2 knockout mice, angiotensin II increased matrix metalloproteinase (MMP)-2 and MMP-9 activity by overexpressed 120% and 390% compared with wild-type mice (P< .05). In conclusion, we identify TSP2 as a crucial regulator of the integrity are of the cardiac matrix that is necessary for the myocardium to cope with increased loading and that may function by it its regulation of MMP activity. This suggests that expression of TSP2 marks an early-stage molecular program that is activated uniquely ( .19+/- .01) in hypertrophied hearts that are prone to fail.

Embryonic expression stem (ES) cells have the ability to differentiate into a variety of cell lineages. We are examining ES cell differentiation knowing in vitro by using cDNA microarrays to generate a molecular phenotype for each cell type. El4 ES cells induced by obtained retinoic acid after forming embryoid bodies differentiate almost exclusively to neurons. We obtained expression patterns for about 8500 gene sequences of by comparing mRNAs from undifferentiated ES cells and their differentiated derivatives in a competitive hybridization. Our results indicate that the Gene genes expressed by ES cells change dramatically as they differentiate (58 gene sequences up-regulated, 34 down-regulated). Most notably, totipotent ES transcriptional cells expressed high levels of a repressor of Hox expression (the polycomb homolog Mphl) and a co-repressor (CTBP2). Expression of ability these genes was undetectable in differentiated cells; the ES cell-derived neurons expressed a different set of transcriptional regulators, as weil into as markers of neurogenesis. The gene expression profiles indicate that ES cells actively suppress differentiation by transcriptional repression; cell-cell contact and in embryoid bodies and retinoic acid treatment may overcome this suppression, allowing expression of Hox genes and inducing a suite Most of neuronal genes. Gene expression profiles will be a useful outcome measure for comparing in vitro treatments of differentiating ES cDNA cells and other stem cells. Also, knowing the molecule phenotype of transplantable cells will allow correlation of phenotype with the differentiate success of the transplant.

Recent and development of gene expression profiling technologies has enabled the large-scale analysis of gene expression changes during disease progression. Frequently, cardiovascular number diseases involve complex interactions of multiple cell types over prolonged periods of time. A better understanding of the pathology of changes cardiovascular diseases and the potential identification of underlying genetic defects are currently being explored by using profiling methodologies in a better number of animal and tissue-culture models.

We pattern conducted large scale gene expression analysis of the response of macrophages to exposure to oxidized low density lipoprotein (Ox-LDL). Much macrophages. of the vessel wall lesion of atherosclerosis is composed of macrophages that have become engorged with cholesterol. These resulting "foam several cells" contribute to the progression of vascular disease through several pathways. As a potential model of foam cell formation, we at treated THP-1 cells with 12-O-tetradecanoylphorbol 13-acetate to differentiate them into a macrophage-like phenotype and subsequently treated them with oxidized low components density lipoprotein for various time periods. RNA from Ox-LDL treated and time-matched control untreated cells was hybridized to microarrays containing profiles. 9808 human genes. 268 genes were found to be at least 2-fold regulated at one or more time points. These analysis regulation patterns were classified into seven clusters of expression profiles. The data is discussed in terms of the overall pattern the of gene expression, the thematic classification of the responding genes, and the clustering of functional groups in distinct expression patterns.groups The magnitude and the temporal patterns of gene expression identified known and novel molecular components of the cellular response that time-matched are implicated in the growth, survival, migratory, inflammatory, and matrix remodeling activity of vessel wall macrophages. In particular, the role wall of nuclear receptors in mediating the gene expression modulation by Ox-LDL is highlighted.

The into ABC1 transporter was identified as the defect in Tangier disease by a combined strategy of gene expression microarray analysis, genetic emphasized mapping, and biochemical studies. Patients with Tangier disease have a defect in cellular cholesterol removal, which results in near zero removal, plasma levels of HDL and in massive tissue deposition of cholesteryl esters. Blocking the expression or activity of ABC1 reduces by apolipoprotein-mediated lipid efflux from cultured cells, and increasing expression of ABC1 enhances it. ABC1 expression is induced by cholesterol loading ABC1 and cAMP treatment and is reduced upon subsequent cholesterol removal by apolipoproteins. The protein is incorporated into the plasma membrane removal in proportion to its level of expression. Different mutations were detected in the ABC1 gene of 3 unrelated patients. Thus,identified ABC1 has the properties of a key protein in the cellular lipid removal pathway, as emphasized by the consequences of the its defect in patients with Tangier disease.